Fig. 4

Ex vivo functional assay of bioengineered intestinal grafts. a Photographs of a cadaveric and a recellularized intestinal scaffold undergoing ex vivo perfusion in a thermostatic chamber. Grafts were perfused with vascular perfusate through the cannulated superior mesenteric artery (SMA), while the intestinal lumen was instilled with luminal perfusate that contained nutritional elements. Vascular effluent was collected through the cannulated superior mesenteric vein (SMV). Scale bars, 1 cm. b Diagram of the isolated perfusion circuit used for ex vivo functional analysis. Oxygenated Krebs–Heinseleit buffer (KHB) was perfused through the SMA under constant pressure of 80 mmHg, while luminal perfusate (L) was infused at a constant rate of 0.6 ml min⁻¹ using a peristaltic pump (P). c Flow rates of vascular perfusate measured by venous effluent collection of cadaveric, decellularized, and regenerated intestines (n = 3), showing loss of vascular flowthrough with decellularization and partial restoration in regenerated intestines. The data were normalized by tissue mass. d Luminal-to-vascular glucose transfer rate of cadaveric, decellularized, and regenerated intestines (n = 3), showing loss of glucose absorptive function after decellularization and nearly-significant restoration in regenerated intestines. The data were normalized by length of intestine. e Luminal-to-vascular 6-Fluorescein-5(6)-carboxamido hexanoic acid (FHA) transfer rates of cadaveric, decellularized, and regenerated intestines (n = 3), showing increased FHA transfer in decellularized compared to cadaveric intestine, but a similar rate to cadaveric tissue in regenerated intestines. The data were normalized by length of intestine. Error bars, mean ± s.d. of experimental values. *P < 0.05; **P < 0.01, Student’s t test. Ab, aboral end of the intestinal lumen; C, carbogen; EC, effluent collection; JC, thermostatic jacketed chamber; O, oral end of the intestinal lumen; S, pressure sensor